[Histonet] peristaltic pump setup/storage of frozen rat brain

From:

Michael Patric

Hello Everyone,
A colleague recommended this group to me. I hope you will be able to help me with three questions that I have:
1) I am trying to setup a Masterflex L/S peristaltic economy drive pump system to assist in the perfusion of adult rats. We are interested in collecting the brain for later immunohistichemistry/in situ studies. I would like to know what the appropriate type of tubing and tube size should be for this setup?
2) I am embedding these fixed brains in OCT for freezing with N2/isopentane. I have been trying to find molds that are of appropriate size for freezing an entire adult rat brain. The commerical molds (Tissue Tek) I have seen are either too big or too small for freezing an adult rat brain en bloc. Does anyone have a recommedation or do they use something completely different? Also is better cut these brains into smaller sections and freezing them rather freezing the entire brain en block?
3) Finally, can someone recommend their procedure for storing rat brains (probably will be the same for any biological tissue) in a -80 freezer for long-term storage?
Appreciate the help,
Michael Patrick
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